Ants moderate the supply of critical resources such as water and nutrients in desert environments by changing the physical arrangement of soils during nest construction. We measured water infiltration and soil physical and chemical properties on and off the nests of two ant species (Pogonomyrmex rugosus, Aphaenogaster cockerelli) across five sites at differing landscape positions within a Chihuahuan Desert watershed. Our aim was to test whether the effects of these long-lived ant nests on water infiltration and soil chemistry varied between ant species or across sites within a watershed. Water flow was generally slowest at the site with the highest silt and clay contents, and fastest at the site with sandy soils. Flow was generally greater through ant nest soils than adjacent non-nest soils, and we attribute this to increases in macropores in the nests. However, the effects of both ant nests and species varied among sites. Despite wide variation in soil chemical properties across all sites, ant nests had a consistent effect on soil chemical properties, with higher levels of carbon, nitrogen, sulphur, phosphorus and electrical conductivity on nests compared with non-nest soils. Our results demonstrate that while we can generalise about the effects of ant nests on water flow and nutrient levels, differences in soil type, nest density and ant species across sites are likely to moderate these effects.